DNA

Part:BBa_K1031021

Designed by: He Shuaixin   Group: iGEM13_Peking   (2013-09-09)
Revision as of 06:24, 25 September 2013 by Sclereid (Talk | contribs)

Hybrid Promoter for Band-pass Filter

hybrid promoter for the band-pass filter


Construction of our hybrid promoter

After deciding the circuit and protein to use, we started to design the promoter that could be activated by phiR73δ and be repressed by cI. To obtain a promoter, a hybrid promoter is required for the reporter node. We modified bacteriophage φR73’s promoter into a hybrid promoter that can be activated by the PhiR73 delta activator and repressed by the repressor cI respectively. We replaced the sequence between the -10 and -35 elements with the cI binding site Or1 from Phage λ. PhiR73δ activator binds to its binding site and the transcription starts. The binding of cI dimers will block the binding of σ70 factors and therefore repress the transcription (Fig.).

Peking2013_Construction_of_hybrid_promoter.png

Figure 5.The construction of our hybrid promoter.

Based on this construction, when the input signal is weak, the concentration of PhiR73 delta is too low to generate a strong output. When the input signal is medium, despite a portion of promoters occupied by cI dimmers, the rest still can be activated by PhiR73 delta and bring about a visible output. When the input signal is strong, almost all of the promoters are “conquered” by cI dimers. The transcription is blocked and the output is shut down. Hence only medium input signal can generate a significant output and the Band Pass Filter was made.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 16
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 127


[edit]
Categories
Parameters
n/aHybrid Promoter for Band-pass Filter